Study On The Pharmacological Activity Of Non-water-soluble Components From Oviductus Ranae

Purpose:1.To investigate the optimal enzymatic technology of NWSC (non-water-soluble components) from Oviductus Ranae.2.To research the chemical composition of NWSC and HNWSC (non-water-soluble components hydrolysates) from Oviductus Ranae.3.To research the pharmacological activity of NWSC and HNWSC from Oviductus Ranae.Method: 1.To explore the optimal enzymatic technology of NWSC from Oviductus RanaeThe viscosity index, by single factor test and L9(34)orthogonal test to explore the effects of enzyme concentration, hydrolysis temperature, hydrolysis time of NWSC from Oviductus Ranae, to establish the optimal enzymatic technology of NWSC from Oviductus Ranae.2.To research the chemical composition of NWSC and HNWSC from Oviductus Ranae.Using SDS-PAGE combine gel imaging technology analyze the molecular weight of Oviductus Ranae non-enzymatic water-soluble components before and after enzyme; using pre-column derivatization-high performance liquid chromatography measure the amino acid content of NWSC and HNWSC from Oviductus Ranae.3.The lipid-lowering effect of NWSC and HNWSC from Oviductus Ranae.Feeding normal rats high fat diet to establish high-fat rats model, the rats were randomly divided into the basal diet control group, model group, NWSC (low, medium and high dose) group and HNWSC (low, medium and high dose) group,intragastric continue for 30 days, the rats measured total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and other content.4.The anti-fatigue effect of NWSC and HNWSC from Oviductus Ranae.Intragastric continue for 30 days with low, middle and high dose of NWSC and HNWSC from Oviductus Ranae, blank control group were given equal volume of distilled water, observed the swimming time, the serum urea nitrogen, hepatic glycogen, blood lactic acid content of mice respectively.5.The anti-hypoxia effect of NWSC and HNWSC from Oviductus Ranae. Set low, middle and high dose groups and blank control group of NWSC and HNWSC from Oviductus Ranae, intragastric continue for 30 days, measured hypoxia tolerance time, the survival time of sodium nitrite poisoning, acute ischemic anoxia survival time of mice.6.The antioxidant effect of NWSC and HNWSC from Oviductus Ranae.Except the control group, all other groups were intraperitoneally injected with D-galactose solution at the daily dose of 100 mg/kg bw·d for 6 successive weeks. The mice were divided into eight groups, intragastric continue for 30 days.Malondialdehyde (MDA)content, glutathione peroxidase (GSH-Px) activity and Super oxide dismutase (SOD) activity in liver were assayed.Result: 1.The study proved out the best hydrolysis condition of NWSC of Oviductus Ranae were: enzyme dosage is 250u/g, reaction temperature is 65℃, hydrolysis time is 60min.2.After hydrolysis,the molecular weight of NWSC of Oviductus Ranae decreased significantly,the hydrolyze process was completely;the content of amino acids in the enzyme did not change significantly.3.NWSC of Oviductus Ranae could significantly reduce the TG content in hyperlipidemic rats (P<0.05); the high dose group decreased the TC content significantly in hyperlipidemic rats (P<0.05),had no significant effect on HDL-C content.HNWSC of Oviductus Ranae could reduce the hyperlipidemic rats TG and TC levels significantly (P <0.05), had no effect on HDL-C level.4. NWSC of Oviductus Ranae in the middle dose group of swimming time was longer than the control group significantly (P<0.05); every dose of glycogen content were higher significantly (P<0.05); the area under the curve of blood lactate in the middle and high dose group was less than the control group (P <0.05); after exercise, blood urea nitrogen in mice of all dose groups showed no significant change.HNWSC of Oviductus Ranae of swimming time in all dose groups compared with the control group in mice was longer significantly (P<0.05), low-dose group compared with the control group of liver glycogen content was increased significantly (P<0.05), the area under the curve of blood lactate in the dose group compared with the control group decreased significantly (P<0.01), all dose groups compared with the control group of serum urea nitrogen had no significant difference (P>0.05).5.All dose group of the hypoxia tolerance survival time, survival time of sodium nitrite poisoning, acute cerebral ischemia and hypoxia survival time of Oviductus Ranae NWSC had no significant difference.HNWSC of Oviductus Ranae in all dose groups in the survival time of hypoxia tolerance had significant difference,the middle and high dose groups in nitrite poisoning survival time had significant difference, the middle dose group in acute cerebral hypoxic-ischemic survival time had significant difference.6. NWSC of Oviductus Ranae exhibited decrease in MDA content,GSH-Px activity revealed a significant increase in the low-dose group.HNWSC of Oviductus Ranae exhibited decrease in MDA content,GSH-Px and SOD activity revealed increase in all dose groups.Conclusion: 1.The HNWSC of Oviductus Ranae from the initial viscosity 2325.50mPa.s reduced to 11.17mPa.s, the mobility and solubility was better. The hydrolysis process is simple, stable and feasible.2.After hydrolysis,the molecular weight of NWSC of Oviductus Ranae decreased significantly;Oviductus Ranae NWSC and HNWSC contain identical amino acids, the content of amino acids in the hydrolyze did not change significantly.3.The NWSC and HNWSC from Oviductus Ranae had assisted lowering blood fat effect, and the effect of HNWSC was further enhanced.4.The NWSC and HNWSC from Oviductus Ranae had anti-fatigue effect, and the effect of HNWSC was further enhanced.5.The NWSC of Oviductus Ranae had no effect on anti-hypoxia, the HNWSC had significant anti-hypoxia effect.6.The NWSC and HNWSC from Oviductus Ranae had antioxidant effect, and the effect of HNWSC was further enhanced.